Biomimetic Extracellular Matrices

Abstract

The present invention is directed to hydrogel compositions for biotechnology applications. Specifically, the invention provides hydrogels mimicking the ECM and uses thereof.

1 . A composition comprising a copolymer comprised of a first and a second dextran macromonomer wherein each of said first and second dextran macromonomer comprises a different degree of glycidylmethacrylate (GMA) substitution. 
     
     
         2 . The composition of  claim 1 , wherein the degree of substitution in the first macromonomer is between about 1/23 and 1/10. 
     
     
         3 . The composition of  claim 1 , wherein the degree of substitution in the second macromonomer is between about 1/23 and 1/10. 
     
     
         4 . The composition of  claim 1 , wherein said substitution is a methacrylate-hydroxyl substitution, or a carbonate-hydroxyl substitution. 
     
     
         5 . The composition of  claim 1 , further comprising an extra cellular matrix protein covalently bound to said dextran macromonomer. 
     
     
         6 . The composition of  claim 5 , wherein said extra cellular matrix protein is laminin, fibronectin, collagen or their combination. 
     
     
         7 . A biomimetic hydrogel comprising a copolymer having a first and a second dextran macromonomer wherein each said dextran macromonomer comprises a different degree of glycidylmethacrylate (GMA) substitution; and crosslinking agent. 
     
     
         8 . The hydrogel of  claim 7 , wherein the degree of substitution in the first macromonomer is between about 1/23 and 1/10. 
     
     
         9 . The hydrogel of  claim 7 , wherein the degree of substitution in the second macromonomer is between about 1/23 and 1/10. 
     
     
         10 . The hydrogel of  claim 7 , wherein the substitution is a methacrylate-hydroxyl substitution, or a carbonate-hydroxyl substitution. 
     
     
         11 . The hydrogel of  claim 7 , further comprising a covalently bound extra cellular matrix protein to the dextran macromonomer. 
     
     
         12 . The hydrogel of  claim 11 , wherein the extra cellular matrix protein is laminin, fibronectin, collagen or their combination. 
     
     
         13 . The hydrogel of  claim 7 , wherein the storage modulus (G′) is between about 400 pa to 42 Kpa. 
     
     
         14 . The hydrogel of  claim 7 , wherein the catalyst is TEMED, DEMED or their combination. 
     
     
         15 . The hydrogel of  claim 14 , wherein further comprising ammonium persulfate (APS), or riboflavin. 
     
     
         16 . A three-dimensional tissue scaffold for supporting tissue on-growth, the scaffold comprising: a substrate immobilized hydrogel, wherein the hydrogel comprises a copolymer having a first and a second dextran macromonomer wherein each of said first and second dextran macromonomer comprises a different degree of glycidylmethacrylate (GMA) substitution; and at least one of a living cell, an ECM ligand, protein, peptide, transcript factor, cytokine, therapeutic agent, growth factor, encapsulated in the hydrogel or on its surface. 
     
     
         17 . The tissue scaffolding of  claim 16 , wherein said substrate comprises cobalt, silicon, plastic, chromium, glass, stainless steel, polystyrene, tantalum, titanium, carbon, calcium, quartz, ceramic, or a combination thereof. 
     
     
         18 . The tissue scaffolding of  claim 16 , wherein the cells are myocyte precursor cells, cardiac myocytes, skeletal myocytes, satellite cells, fibroblasts, cardiac fibroblasts, chondrocytes, osteoblasts, endothelial cells, epithelial cells, embryonic stem cells, hematopoetic stem cells, neuronal cells, mesenchymal stem cells, anchorage-dependent cell precursors, or combinations thereof. 
     
     
         19 . The tissue scaffolding of  claim 16 , further comprising tissue grown over the scaffolding. 
     
     
         20 . A method of making an implant for supporting tissue on-growth, the method comprising: providing the three-dimensional tissue scaffold of  claim 16 . 
     
     
         21 . A preconditioned artificial tissue comprising living cells that are attached to the three-dimensional tissue scaffold of  claim 16 . 
     
     
         22 . The scaffold of  claim 16 , wherein the degree of substitution in the first macromonomer is between about 1/23 and 1/10. 
     
     
         23 . The scaffold of  claim 16 , wherein the degree of substitution in the second macromonomer is between about 1/23 and 1/10. 
     
     
         24 . The scaffold of  claim 16 , wherein the substitution is a methacrylate-hydroxyl substitution, or a carbonate-hydroxyl substitution. 
     
     
         25 . The scaffold of  claim 16 , further comprising a covalently bound extra cellular matrix protein to the dextran macromonomer. 
     
     
         26 . The scaffold of  claim 26 , wherein the extra cellular matrix protein is laminin, fibronectin, collagen or their combination. 
     
     
         27 . The scaffold of  claim 16 , wherein the storage modulus (G′) is between about 400 pa to 42 Kpa. 
     
     
         28 . The scaffold of  claim 27 , whereby the storage modulus is no more than 1 Kpa. 
     
     
         29 . The scaffold of  claim 16 , whereby the living cell maintains its phenotypic structure. 
     
     
         30 . An implant comprising the scaffold of  claim 16 . 
     
     
         31 . A method of modulating the amount and location of an ECM ligand attachment on the surface of a hydrogel comprising the step of: selectively modifying the aldehyde concentration and location at the surface of a hydrogel comprising a first and a second dextran macromonomer wherein each of said first and second dextran macromonomer comprises a different degree of glycidylmethacrylate (GMA) substitution; and covalently binding the ECM ligand to the aldehyde group at the surface of the hydrogel. 
     
     
         32 . The method of  claim 31 , whereby the step of selectively modifying the aldehyde concentration at the surface of the hydrogel comprises exposing only the surface of the hydrogel to periodate oxidation. 
     
     
         33 . The method of  claim 31 , whereby the periodic acid is immobilized on a substrate. 
     
     
         34 . The method of  claim 31 , whereby the step of covalently binding the ECM ligand to the aldehyde group at the surface of the hydrogel comprises the step of contacting the surface modified hydrogel with the ECM ligand 
     
     
         35 . The method of  claim 31 , whereby the ECM ligand is laminin, fibronectin, collagen or their combination. 
     
     
         36 . The method of  claim 31 , whereby the degree of substitution in the first macromonomer is between about 1/23 and 1/10. 
     
     
         37 . The method of  claim 31 , whereby the degree of substitution in the second macromonomer is between about 1/23 and 1/10. 
     
     
         38 . The method of  claim 31 , whereby the substitution is a methacrylate-hydroxyl substitution, a carbonate-hydroxyl substitution, or a mixture thereof. 
     
     
         39 . The method of  claim 31 , whereby said substrate comprises cobalt, silicon, plastic, chromium, glass, stainless steel, polystyrene, tantalum, titanium, carbon, calcium, quartz, ceramic, or a combination thereof. 
     
     
         40 . A method of culturing living cells in a hydrogel while maintaining their phenotypic structure, comprising the steps of: encapsulating the cells in a hydrogel comprising a copolymer comprised of a first and a second dextran macromonomer wherein each of said first and second dextran macromonomer comprises a different degree of glycidylmethacrylate (GMA) substitution; modulating the storage modulus of the hydrogel; covalently binding the hydrogel to an ECM ligand; and allowing the cells to grow. 
     
     
         41 . A method of controlling cell culture proliferation on a hydrogel surface, comprising the steps of immobilizing a hydrogel comprising a copolymer of a first and a second dextran macromonomer wherein each of said first and second dextran macromonomers comprises a different degree of glycidylmethacrylate (GMA) substitution, onto a substrate; modulating the storage modulus of the immobilized hydrogel; functionalizing the surface of the hydrogel with an ECM ligand; and seeding the hydrogel with the cell culture whose proliferation is sought to be controlled, wherein below a threshold storage modulus, no cell proliferation will occur. 
     
     
         42 . The method of  claim 40 , whereby each of the first and second macromonomer in the copolymer has a degree of substitution of between about 1/23 and 1/10. 
     
     
         43 . The method of  claim 42 , whereby the substituted dextran macromonomer is a methacrylate-hydroxyl substituted dextran macromonomer, a carbonate-hydroxyl substituted dextran macromonomer, or a mixture thereof. 
     
     
         44 . The method of  claim 42 , whereby the step of immobilizing the hydrogel onto a substrate comprises the steps of:
 (a) activating the substrate;   (b) immersing the activated substrate in an agent capable of forming a covalent bond between said activated substrate and said composition thereby making a functionalized substrate;   (c) drying said functionalized substrate, and   (d) contacting the functionalized substrate with the hydrogel   
     
     
         45 . The method of  claim 42 , whereby said substrate comprises cobalt, silicon, plastic, chromium, glass, stainless steel, polystyrene, tantalum, titanium, carbon, calcium, quartz, ceramic, or a combination thereof.